RNA Detection and Visualization

Volume 714 of the series Methods in Molecular Biology pp 407-422


RIP: An mRNA Localization Technique

  • Sabarinath Jayaseelan
  • , Francis Doyle
  • , Salvatore Currenti
  • , Scott A. TenenbaumAffiliated withDepartment of Nanobioscience, University at Albany Email author 

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A detailed understanding of post-transcriptional gene expression is necessary to correlate the different elements involved in the many levels of RNA–protein interactions that are needed to coordinate the cellular biomolecular machinery. The profile of mRNA, a major component of this machinery, can be examined after isolation from specific RNA-binding proteins (RBPs). RIP-Chip or ribonomic profiling is a versatilein vivo technique that has been widely used to study post-transcriptional gene regulation and the localization of mRNA. Here we elaborately detail the methodology for mRNA isolation using RBP immunoprecipitation (RIP) as a primary approach. Specific antibodies are used to target RBPs, which are then used to capture the associated mRNA.

Key words

Ribonomics mRNA localization RNA-binding protein RIP-Chip RNA coprecipitation